FastDigest Pfl23II

Protocol tips

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	Four-fifth of each PCR product were purified (Qiagen PCR purification kit) and digested with 1 μl of FastDigest Pfl23II (Thermo Fisher Scientific) for 1 hour at 37 °C. After heat inactivation at 65 °C for 15 min, the digested products were analysed on a 2% agarose gel as well as the remainder of each undigested PCR product (Supplementary Fig. S5).

Protocol tips
Four-fifth of each PCR product were purified (Qiagen PCR purification kit) and digested with 1 μl of FastDigest Pfl23II (Thermo Fisher Scientific) for 1 hour at 37 °C. After heat inactivation at 65 °C for 15 min, the digested products were analysed on a 2% agarose gel as well as the remainder of each undigested PCR product (Supplementary Fig. S5).

Publication protocol

"rDNA analysis
Genomic DNA was isolated from individual female mosquitoes originated from crosses between CRISPRSD males and wild-type females using the Wizard Genomic DNA Purification Kit (Promega). Genomic loci containing the CRISPRSD target sites were amplified with Phusion HF polymerase (Thermo Scientific) using RG236-GCAGCATAGGTGGGAGGGCTTCCTC and RG238-GTTTCTGTCCACACTGAGCTGACCT primers. Four-fifth of each PCR product were purified (Qiagen PCR purification kit) and digested with 1 μl of FastDigest Pfl23II (Thermo Fisher Scientific) for 1 hour at 37 °C. After heat inactivation at 65 °C for 15 min, the digested products were analysed on a 2% agarose gel as well as the remainder of each undigested PCR product"

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Manufacturer protocol

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